Name: _____________________________________ Class: ________________ Date: ____________

Effect of Temperature on Amylase

Amylase is an enzyme that catalyzes the breakdown of starch, a polysaccharide, into maltose, a disaccharide. This reaction is called hydrolysis, because a molecule of water is needed to break each bond.

Amylase is naturally produced as part of saliva, and released into the oral cavity. As an enzyme, amylase is sensitive to many environmental factors, such as temperature and pH. The purpose of this lab is to determine the optimal temperature for amylase to catalyze the hydrolysis of starch.

Materials
0.1% starch solution (ice bath, room temperature, body temperature, and boiling) Six test tubes Graduated cylinder Digital Timer

Pre-Lab Questions
1. Starch is a polysaccharide, while maltose is a disaccharide. What is the difference between the two?

2. What kind of molecule are enzymes? (Carbohydrate, lipid, protein, or nucleic acid)

3. In general, what is the purpose of enzymes in the human body?

4. Explain what denaturation is, and how it specifically affects enzymes.

5. Which chemical test (Benedicts, Biuret, Iodine, Sudan) is effective for starch? For smaller sugars?

6. Hypothesis: Given what you know about the human digestive system, what temperature do you predict will be ideal for hydrolysis of starch by amylase enzyme?

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Name: _____________________________________ Class: ________________ Date: ____________

Procedure
7. Measure 5mL of each temperature of starch solution and place it into four separate test tubes. Make sure the test tubes are marked or labeled to tell them apart. 8. Add 2 drops of iodine to each test tube. 9. Add 2mL of amylase enzyme to each test tube. Start the timer. 10. Place the test tubes back in the same temperature that you got the starch solutions from. For example, the test tube filled from the ice bath starch should go back into an ice bath. 11. As a negative control, measure a test tube with 5ml of water. Add iodine. 12. As a positive control, measure a test tube with 5mL of starch solution (any temperature). Add iodine. 13. Record on the data table how long it takes for each test tube to change color. When the starch is hydrolyzed, it should look like the negative control iodine (yellowish-brown). Note: If the starch has not been hydrolyzed after 10 minutes, move onto the next reaction.

Data
Table 1 - Effects of Temperature on Hydrolysis Reaction Length (Individual Data)

Temperature (C) Ice Bath Room Temperature Warm Bath Boiling

Reaction Length (s)

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Name: _____________________________________ Class: ________________ Date: ____________

Table 2 - Effects of Temperature on Hydrolysis Reaction Length (Class Data)

Ice Bath

Reaction Length (s) Room Warm Bath Temperature

Boiling

Table 3 - Class Average Hydrolysis Reaction Time

Ice Bath

Reaction Length (s) Room Warm Bath Temperature

Boiling

Figure 1- Graph of Enzyme Temperature and Average Reaction Time

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Name: _____________________________________ Class: ________________ Date: ____________

Post-Lab Questions
14. Conclusion: Do the results support your hypothesis? Be specific.

15. Why was iodine used to monitor the starch, instead of using Benedicts solution to monitor the appearance of simple sugars?

16. Was there any evidence of enzyme denaturation in any of the trials?

17. Look at the table of class data. Were there any results that might be considered outliers? What are some possible sources of error that could cause an outlier?

18. One other possible variable that influences the action of enzymes like amylase is pH level. What pH would you expect to be optimal for the action of this enzyme? Test this idea by taking a sample of saliva and adding it to a pH strip.

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